Functional display of foreign protein on surface of Escherichia coli using N-terminal domain of ice nucleation protein
Functional display of foreign protein on surface of Escherichia coli using N-terminal domain of ice nucleation protein
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DOI：
10.1002/bit.10892
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发表时间：
2004-01-20
影响因子：
3.8
通讯作者：
Cha, HJ
中科院分区：
文献类型：
--
作者：
Li, L;Kang, DG;Cha, HJ
我们研究了InaK的N-末端结构域的能力，InaK是一种来自假单胞菌KCTC 1832的冰核蛋白，作为在大肠杆菌细胞表面展示外源蛋白的锚定基序。将绿色荧光蛋白（GFP）与InaK的N-末端结构域（InaKN）或已知的含有N-和C-末端结构域的截短的InaK（InaK-NC）融合后，比较GFP的总表达水平和表面展示效率。我们报道了InaK-N/ GFP融合蛋白显示出与InaK-NC/GFP相似的细胞表面展示效率（类似于50%），表明单独的InaK N-末端区域可以指导外源蛋白易位到细胞表面，并且可以用作潜在的细胞表面展示基序。此外，基于单位细胞密度，InaK-N/GFP显示出最高水平的总表达和表面展示。InaK-N还成功地指导了有机磷水解酶（OPH）的细胞表面展示，证实了其作为展示基序的能力。(C)2004 Wiley Periodicals，Inc.
We investigated the ability of the N-terminal domain of InaK, an ice nucleation protein from Pseudomonas syringae KCTC 1832, to act as an anchoring motif for the display of foreign proteins on the Escherichia coli cell surface. Total expression level and surface display efficiency of green fluorescent protein (GFP) was compared following their fusion with either the N-terminal domain of InaK (InaKN), or with the known truncated InaK containing both N- and C-terminal domains (InaK-NC). We report that the InaK-N/ GFP fusion protein showed a similar cell surface display efficiency (similar to 50%) as InaK-NC/GFP, demonstrating that the InaK N-terminal region alone can direct translocation of foreign proteins to the cell surface and can be employed as a potential cell surface display motif. Moreover, InaK-N/GFP showed the highest levels of total expression and surface display based on unit cell density. InaK-N was also successful in directing cell surface display of organophosphorus hydrolase (OPH), confirming its ability to act as a display motif. (C) 2004 Wiley Periodicals, Inc.